• Journal of Mushroom
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• v.10 no.3
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• pp.143-149
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• 2012

Armillaria spp are well known as a symbiotic fungus with Gastrodia elata. This study was carried out to identify and analyze the genetic relationships among 83 strains of Armillaria spp.. The amplified internal transcribed spacer(ITS) region of the rDNA was about 500~750 bp long and identified by 9 strains; A. mellea, A. tabescens, A. ostoyae, A. gallica, A. novae-zenlandia, A. cepistipes, A. nabsnona, A. gemina, A. sinapina. Sequence analysis showed that 52% of strains were different with original identification. A. gallica, A. cepistipes and A. gemina were so close phylogenetic relationship, that was difficult to classify using ITS region. In A. gallica, 12 strains including ASI10104 were showed a close phylogenetic relationship with A. gallica, A. cepistipes and A. gemina. ASI10017 and ASI10114 were classified as the A. sinapina group, ASI10045 was the A. borealis group, ASI10002 and ASI10025 were the A. ostoyae group. So more studies need for more accurate identification and determine the phylogenetic relationships of Armillaria spp.

• Journal of Life Science
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• v.23 no.1
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• pp.95-101
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• 2013

In this study, plant samples of five species were collected from the Dokdo islands in South Korea. Plant samples such as Asparagus schoberioides, Corydalis platycarpa, Festuca rubra, Sedum oryzifolium, and Setaria viridis were collected from the Dongdo and Seodo. Endophytic fungal strains were isolated from the roots of five plants from the Dokdo islands. Thirty-three fungal strains were isolated from these native plants. All the endophytic fungi were analyzed by internal transcribed spacer (ITS) sequencing (ITS containing ITS1, 5.8s, and the ITS2 region). Waito-c rice seedlings were treated with fungal culture filtrates to test their plant growth-promoting activity. A bioassay of the D-So-1-1 fungal strain isolated from S. oryzifolium confirmed that it has the highest plant growth-promoting activity. All the endophytic fungi belong to four orders: Eurotiales (86%), Capnodiales (3%), Hypocreales (4%), and Incertae sedis (7%). The endophytic fungi were classified as Ascomycota, which contained Aspergillus (12%), Cladosporium (3%), Eurotium (3%), Fusarium (18%), Microsphaeropsis (6%), and Penicillium (58%) at the genus level.

• ALGAE
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• v.29 no.2
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• pp.75-99
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• 2014

A small dinoflagellate, Ansanella granifera gen. et sp. nov., was isolated from estuarine and marine waters, and examined by light microscopy, scanning electron microscopy, and transmission electron microscopy. In addition, the identity of the sequences (3,663-bp product) of the small subunit (SSU), internal transcribed spacer (ITS) region (ITS1, 5.8S, ITS2), and D1-D3 large subunit (LSU) rDNA were determined. This newly isolated, thin-walled dinoflagellate has a type E eyespot and a single elongated apical vesicle, and it is closely related to species belonging to the family Suessiaceae. A. granifera has 10-14 horizontal rows of amphiesmal vesicles, comparable to Biecheleria spp. and Biecheleriopsis adriatica, but greater in number than in other species of the family Suessiaceae. Unlike Biecheleria spp. and B. adriatica, A. granifera has grana-like thylakoids. Further, A. granifera lacks a nuclear fibrous connective, which is present in B. adriatica. B. adriatica and A. granifera also show a morphological difference in the shape of the margin of the cingulum. In A. granifera, the cingular margin formed a zigzag line, and in B. adriatica a straight line, especially on the dorsal side of the cell. The episome is conical with a round apex, whereas the hyposome is trapezoidal. Cells growing photosynthetically are $10.0-15.0{\mu}m$ long and $8.5-12.4{\mu}m$ wide. The cingulum is descending, the two ends displaced about its own width. Cells of A. granifera contain 5-8 peripheral chloroplasts, stalked pyrenoids, and a pusule system, but lack nuclear envelope chambers, a nuclear fibrous connective, lamellar body, rhizocysts, and a peduncle. The main accessory pigment is peridinin. The SSU, ITS regions, and D1-D3 LSU rDNA sequences differ by 1.2-7.4%, >8.8%, and >2.5%, respectively, from those of the other known genera in the order Suessiales. Moreover, the SSU rDNA sequence differed by 1-2% from that of the three most closely related species, Polarella glacialis, Pelagodinium bei, and Protodinium simplex. In addition, the ITS1-5.8S-ITS2 rDNA sequence differed by 16-19% from that of the three most closely related species, Gymnodinium corii, Pr. simplex, and Pel. bei, and the LSU rDNA sequence differed by 3-4% from that of the three most closely related species, Protodinium sp. CCMP419, B. adriatica, and Gymnodinium sp. CCMP425. A. granifera had a 51-base pair fragment in domain D2 of the large subunit of ribosomal DNA, which is absent in the genus Biecheleria. In the phylogenetic tree based on the SSU and LSU sequences, A. granifera is located in the large clade of the family Suessiaceae, but it forms an independent clade.

• Microbiology and Biotechnology Letters
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• v.39 no.4
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• pp.324-329
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• 2011

Eight endophytic fungal strains were isolated from the roots of Calystegia soldanella from the western coast of South Korea. The culture filtrate of the eight endophytic fungi were applied to waito-c rice seedlings in order to verify potential plant growth promotion activities. The results of bioassay indicated that the Cs-9-7 fungal strain possessed the highest plant growth promotion activity. Fungal culture filtrates were analyzed to verify secondary metabolites using gas chromatography and mass spectroscopy with selected ion monitoring (GC/MS-SIM). The culture filtrate of the Cs-9-7 fungal strain was confirmed to contain gibberellins GA3 (1.229 ng/mL), GA4 (3.535 ng/mL), GA7 (1.408 ng/mL) and GA12 (0.378 ng/mL). Polymerase chain reactions (PCR) were performed so as to determine the internal transcribed spacer (ITS) regions for the identification of isolated strains with universal primers ITS-1 and ITS-4. The Cs-9-7 fungal strain, isolated from the root of C. soldanella, has been named Aspergillus tubingensis Cs-9-7.

• Microbiology and Biotechnology Letters
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• v.40 no.4
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• pp.287-295
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• 2012

Eighty-four endophytic fungal strains were isolated and identified from the roots of halophytes collected in Buan salt marsh. All halophyte samples, such as Suaeda japonica, and Suaeda maritima were isolated from Buan salt marsh. All endophytic fungi isolated were analyzed by sequences of internal transcribed spacer (ITS) containing ITS1, 5.8s and ITS2 region. All endophytic fungi expressed that fungal strains belong to eight orders; Pleosporales (45%), Eurotiales (27%), Incertae sedis (11%), Dothideales (6%), Capnodiales (5%), Hypocreales (5%), and Agaricales (1%). All endophytic fungi were confirmed at the genus level of Ascomycota and Basidiomycota, containing Alternaria, Ascomycota, Aspergillus, Aureobasidium, Cladosporium, Eupenicillium, Fusarium, Gibberella, Hypocrea, Lewia, Macrophoma, Penicillium, Peyronellaea, Phoma, Pleospora, Pleosporales, Pseudeurotium, Schizophyllum, and Talaromyces. Alternaria (21%) and Penicillium (13%) were the dominant endophytic fungal strains. In this study, endophytic fungal strains analyzed from S. japonica and S. maritime, Alternaria (21%), and Penicillium (13%) of Pleosporales and Eurotiales in halophytes were very abundant.

• Journal of Life Science
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• v.19 no.3
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• pp.311-316
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• 2009

The brown algae, or phaeophytes, are a large group of multicellular algae, including many notable types of seaweed. We analysed intra- and interspecific phylogenic studies within the genus Pelvetia in Korea and compared them with results of both same and different species in GenBank. The sequences for P. babingtonii in Korea were generally similar to those for P. babingtonii AF102957, and the sequences of P. siliquosa in Korea were also similar to those of P. siliquosa AF102958. Sequence variation within the Pelvetia is mostly due to nucleotide substitutions, although several small indels and some large indels can be found. Another source of sequence divergence is length variation due to stretches of short repeats that occur at the ITS1 or ITS2 in all the Pelvetia. NJ analysis consists mainly of two clades. One of them contains P. canaliculata and P. limitata, and is sister to the rest of the genus (P. siliquosa, P. compressa, and P. babingtonii). P. babingtonii is not grouped one clade. In the MP analysis, ten accessions or populations were fully resolved and all accessions from the same species formed with 99% or 100% bootstrap supports.

• The Korean Journal of Mycology
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• v.37 no.1
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• pp.86-90
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• 2009

The C. militaris belongs to entomopathogenic fungi, which have their specific sequences in internal transcribed spacer regions (ITS1 and ITS2) depending on species. In this study, to identify the phylogenetic relationship of the strain hybrided by mating of C. militaris, we compared DNA sequences of ITS regions and 5.8S ribosomal DNA (rDNA) repeat unit of hybrid strain and its parental strains. The result revealed that hybrid strains are C. militaris species. In addition, cordycepins produced by hybrid strains and other strains of C. militaris were analyzed by HPLC with 20mM $KH_2PO_4$ of mobile phase and C-18 columns. The result indicated that the strain hybrided by mating produce higher concentration of phytochemical cordycepin than other C. militaris strains.

• Journal of Life Science
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• v.22 no.12
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• pp.1718-1723
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• 2012

Ten endophytic fungi with different colony morphologies were isolated from the roots of Suaeda japonica Makino growing naturally in Suncheon Bay. Plant growth promotion was verified by treating waito-c rice seedlings with culture filtrates from the endophytic fungi. The bioassays showed that the Sj/7/4 fungal strain induced effective growth promotion in the seedlings. The gibberellins (GA) produced by fungal strain Sj/7/4 were analyzed by gas chromatography /mass spectroscopy with selected ion monitoring (GC/MS SIM). The culture filtrate of Sj/7/4 fungal strain was confirmed to contain $GA_4$ through quantitative analysis. The Sj/7/4 fungal strain was identified to determine the internal transcribed spacer (ITS) regions with universal primers ITS-1 and ITS-4 by using polymerase chain reactions (PCR). Molecular analysis of the Sj/7/4 fungal strain showed high similarity to Fusarium solani. The Sj/7/4 fungal strain isolated from the root of S. japonica was therefore designated as F. solani Sj/7/4.

• Korean Journal of Plant Taxonomy
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• v.35 no.1
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• pp.7-23
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• 2005

Molecular phylogenetic studies were conducted to evaluate interspecific relationships in 40 populations of Viola including 35 Korean taxa, four Japanese populations and one outgroup using nuclear ribosomal ITS sequences. The phylogenetic analyses were conducted using parsimony and neighbor-joining methods. Subsection Trigonocarpae of section Nomimium appeared as the most basal clade within the Korean Viola. Section Dischidium and Chamaemelanium was monophyletidbootstrap 100%) and placed between subsect. Trigonocarpae and three other subsections of sect. Nomimium. Sect. Nomimium was paraphyletic. Although each subsectional grouping was in accordance with previous infrageneric classification based on morphological characters, yet discordance remained at the series level. Two evolutionary trends observed in the ITS tree were as follows. First, subsect. Trigonocarpae(x=10) was derived from the outgroup(x=6); Second, subsects. Bilobatae and Vaginatae(x=10 or 12), and subsect. Patellares(x=12) of sect. Nomimium were originated from sects. Dischidium and Chamaemelanium(x=6). Viola albida complex including three very closely related taxa was recognized as independent group within subsect. Patellares in parsimony tree. This result suggested that they should be treated as a taxa in series Pinnatae. Phylogenetic position of a putative hybrid species, Viola woosanensis was not supported with previous morphological hypothesis.

• Parasites, Hosts and Diseases
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• v.57 no.1
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• pp.9-15
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• 2019

Melting temperature shift ($T_m-shift$) is a new detection method that analyze the melting curve on real-time PCR thermocycler using SYBR Green I fluorescent dye. To establish a $T_m-shift$ method for the detection of Ancylostoma ceylanicum and A. tubaeforme in cats, specific primers, with GC tail of unequal length attached to their 5' end, were designed based on 2 SNP loci (ITS101 and ITS296) of the internal transcribed spacer 1 (ITS1) sequences. The standard curve of $T_m-shift$ was established using the standard plasmids of A. ceylanicum (AceP) and A. tubaeforme (AtuP). The $T_m-shift$ method stability, sensitivity, and accuracy were tested with reference to the standard curve, and clinical fecal samples were also examined. The results demonstrated that the 2 sets of primers based on the 2 SNPs could accurately distinguish between A. ceylanicum and A. tubaeforme. The coefficient of variation (CV) of $T_m$- values of AceP and AtuP was 0.07% and 0.06% in ITS101 and was 0.06% and 0.08% in ITS296, respectively. The minimum detectable DNA concentration was $5.22{\times}10^{-6}$ and $5.28{\times}10^{-6}ng/{\mu}l$ samples of AceP and AtuP, respectively. The accuracy of $T_m-shift$ method reached 100% based on examination of 10 hookworm DNA samples with known species. In the clinical detection of hookworm in 69 stray cat fecal sample, the $T_m-shift$ detection results were consistent with the microscopic examination and successfully differentiated between the 2-hookworm species. In conclusion, the developed method is a rapid, sensitive and accurate technique and can provide a promising tool for clinical detection and epidemiological investigation of cat-derived hookworms.